tissue culture

From: Kamikaze (011114@hillstrath.on.ca)
Date: Thu Feb 19 1998 - 09:36:00 PST


Date:          Thu, 19 Feb 1998 13:36:00 -0400
From: "Kamikaze" <011114@hillstrath.on.ca>
To: cp@opus.hpl.hp.com
Message-Id: <aabcdefg662$foo@default>
Subject:       tissue culture

Dear list,

I have tried tissue culturing many varities of CP...especially VFt
with some white bulb material attached. When culturing, I sterilized
the plant material in 15% bleach solution for 10 minutes. I then
carefully removed the cuttings into an aquarium that had been filled
with rags soaked in isopropyl alcohol and left inverted on the rags
overnight...in short the aquarium was quite sterile. The media I used
was also well sterilized and did not have any fungal or bacterial
contamination. I then carefully transplanted the sterile (so I
thought) VFT into the TC media inside the aquarium. The VFT culture
after 4 days was contaminated with fungus. I repeated this experiment
several time with varying strengths of bleach and alcohol. In each
case, I either failed to sterilize properly or over sterilized (thus
killing the plant material) the plants. I am at wits end.

Please let me know any experiences that people have had with
effective sterilization agents. I know that the media and aquarium
were quite sterile.

adwait kulkarni



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